抑制miR-193a-5p表达对急性呼吸窘迫综合征大鼠肺纤维化的改善作用及其机制  

Improvement effect of inhibiting miR-193a-5p expression on pulmonary fibrosis in rats with acute respiratory distress syndrome and its mechanism

作　　者：龙光文[1] 张谦[1] 杨秀林[1] 孙鸿鹏[1] 吉春玲[1] LONG Guangwen;ZHANG Qian;YANG Xiulin;SUN Hongpeng;JI Chunling(Department of Emergency,People’s Hospital,Guizhou Province,Guiyang 550002,China)

机构地区：[1]贵州省人民医院急诊内科,贵州贵阳550002

出　　处：《吉林大学学报（医学版）》2024年第6期1491-1498,共8页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金项目(82160021)。

摘　　要：目的:探讨抑制微小RNA(miR)-193a-5p表达对急性呼吸窘迫综合征(ARDS)大鼠肺纤维化的影响,并阐明相关作用机制。方法:60只雄性SD大鼠分为假手术组、模型组、miR-193a-5p拮抗剂组(Antagomir组)和阴性对照组(Antagomir-NC组),每组15只。采用暴露气管法滴注10 mg·kg^(-1)脂多糖(LPS)诱导构建ARDS大鼠模型,假手术组大鼠滴注等体积生理盐水,造模成功后Antagomir组和Antagomir-NC组大鼠给予miR-193a-5p Antagomir或Antagomir-NC尾静脉注射。测定各组大鼠动脉血氧分压(PaO_(2))和氧合指数(OI),HE和Masson染色观察各组大鼠肺组织病理形态表现及肺组织中胶原纤维沉积情况,试剂盒检测各组大鼠肺组织中羟脯氨酸(Hyp)水平,酶联免疫吸附试验(ELISA)法检测各组大鼠肺泡灌洗液(BALF)中炎性因子肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β和IL-6水平,实时荧光定量PCR(RT-qPCR)法检测各组大鼠肺组织中miR-193a-5p表达水平,Western blotting法检测各组大鼠肺组织中β连环蛋白(β-catenin)、蜗牛家族转录抑制因子1(Snail1)和α-平滑肌肌动蛋白(α-SMA)蛋白表达水平。结果:与假手术组比较,模型组大鼠PaO_(2)和OI均明显降低(P<0.05);与模型组比较,Antagomir组大鼠PaO_(2)和OI均明显升高(P<0.05)。HE染色观察,假手术组大鼠肺组织结构正常,未见明显的炎性改变;与假手术组比较,模型组和Antagomir-NC组大鼠肺组织结构轻度异常,肺泡萎缩塌陷,肺泡腔内发现有大量的淋巴细胞和少量的中性粒细胞浸润,肺泡间隙增宽;与模型组比较,Antagomir组大鼠肺组织的肺泡腔内淋巴细胞明显减少,中性粒细胞浸润较少,未见明显增生。Masson染色观察,假手术组大鼠肺组织无明显蓝色胶原纤维沉积;与假手术组比较,模型组和Antagomir-NC组大鼠肺组织中出现大量蓝色胶原纤维沉积,肺泡结构损坏严重,呈现明显的肺纤维化情况;与模型组比较,Antagomir组大鼠肺组织黏液中蓝色Objective:To discuss the effect of inhibiting microRNA(miR)-193a-5p expression on pulmonary fibrosis in the rats with acute respiratory distress syndrome(ARDS),and to clarify the related mechanism.Methods:A total of 60 male SD rats were divided into sham operation group,model group,miR-193a-5p antagonist group(Antagomir group),and negative control group(Antagomir-NC group),and there were 15 rats in each group.The ARDS animal model was induced by administering 10 mg·kg^(-1)lipopolysaccharide(LPS)via tracheal instillation,while the rats in sham operation group received an equal volume of saline.After successful modeling,the rats in Antagomir group and Antagomir-NC group were treated with miR-193a-5p Antagomir or Antagomir-NC via tail vein injection.The arterial partial pressure of oxygen(PaO_(2))and oxygenation index(OI)of the rats in various groups were measured;HE staining and Masson staining were used to observe the pathology and collagen fiber deposition in lung tissue of the rats;kit was used to detect the level of hydroxyproline(Hyp)in lung tissue of the rats in various groups;enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of inflammatory factors tumor necrosis factorα(TNF-α),interleukin(IL)-1β,and IL-6 in bronchoalveolar lavage fluid(BALF)of the rats in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-193a-5p in lung tissue of the rats in various groups;Western blotting method was used to detect the expression levels ofβ-catenin,Snail family transcriptional repressor 1(Snail1),andα-smooth muscle actin(α-SMA)proteins in lung tissue of the rats in various groups.Results:Compared with sham operation group,the PaO_(2)and OI of the rats in model group were significantly decreased(P<0.05);compared with model group,the PaO_(2)and OI of the rats in Antagomir group were significantly increased(P<0.05).The HE staining results showed that the lung tissue structure of the rats in sham operation group was normal,and

关 键 词：微小RNA-193a-5p 急性呼吸窘迫综合征 肺纤维化 WNT/Β-CATENIN信号通路 

分 类 号：R563.9[医药卫生—呼吸系统]